Newcastle disease virus (NDV) infects at least 241 species of pet and free-living birds in addition to domesticated avian species. Wild, feral and domesticated birds are recognized reservoirs of NDV, and contribute to the epidemiology of NDV in the domesticated poultry. The biological and molecular characterization of velogenic NDV (vNDV) from emus is limited. In this study, 54 tissues were collected from eight Emu flocks between May 2010 and January 2012 from highly poultry-dense areas of India including Namakkal, Hyderabad and Bareilly regions. The presence of vNDV was confirmed by virus isolation, fusion (F)-gene based RT-PCR, sequencing of the cleavage site and the virulence scored. One out of eight flocks received from Hyderabad region was found positive for NDV and the in vivo pathotyping revealed the isolate to be vNDV type. The sequence analysis revealed that the isolate had cleavage site of 112-R-R-R-K-R-F-117, which is typical for vNDV. Sequence and phylogenetic analysis of the partial F gene coding regions suggested that the NDV strain belongs to genotype XIII. The Emu isolate had 98100 % nucleotide identity with the vNDVs previously reported in poultry flocks of India. The findings of present study based on the biological and molecular characterization of Emu-origin vNDV, highlights the circulation of genotype XIII in Emus for the first time in the country. There is need to understand the possible spill over of these genetically diverse NDV strains into the commercial poultry and their possible implications in disease control strategies.